Pattern recognition receptors (PRRs) can recognize microbial-specific pathogen-associated molecular patterns, initiate signal cascade transduction, activate the expressions of host immunity and proinflammatory genes, and, ultimately, trigger an immune response against identified pathogens. The present study focused on two outcomes of feeding pearl gentian groupers with high levels of soybean meal (SBM): (1) growth performance and (2) the intestinal environment, including tissue structure, flora profile, and immune responses. Some 720 groupers were randomly divided into three groups (n = 4): (1) controls, fed a 50% fish meal feed (FM), (2) with 20% of the FM substituted with SBM (SBM20), and (3) 40% of the FM substituted with SBM (SBM40). The fish were fed these iso-nitrogenous and iso-lipidic diets for 10 weeks. They were kept in containers with 1 m3 of water under natural light and temperature levels. The experimental results demonstrate that the SBM diets significantly degraded growth performance and intestinal physiology. Typical enteritis characteristics and immune fluctuations appeared, as reflected by the enzyme activities of total superoxide dismutase and lysozyme, and the contents of immunoglobulin M, complement 3, and complement 4. 16SrDNA high-throughput sequencing showed that the intestinal flora was significantly affected, with the abundance of harmful bacteria, such as Vibrio and Streptococcus, increasing with dietary SBM level. Based on “3 + 2” full-length transcriptome sequencing, three triggered PRRs were found in the intestine: the RIG-like receptor, NOD-like receptor, and Toll-like receptor signaling pathways. The intestinal flora variations were significantly correlated with the activation of the three PRR signaling pathways by canonical correlation analysis. These culminated in the transcriptome activation of NF-κB, IRFs, and costimulatory molecules, ultimately promoting the expressions of proinflammatory cytokines, interferons (IFNs), chemokines, and other molecules vital to the innate and/or adaptive immune responses. This study provides new information for diagnosing and preventing SBMIE in aquaculture fish.

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In this study, we present data from an eight-week growth trial with pearl gentian grouper fed either a reference diet (FM) with a fishmeal level of 50%, or test diet wherein 15% (CAP15), 30% (CAP30), 45% (CAP45), and 60% (CAP60) fishmeal was replaced by Clostridium autoethanogenum protein meal (CAP). Results showed that the weight gain and daily feed intake ratio of CAP60 were significantly lower than the FM group. In the serum, compared to the FM group, the content of malondialdehyde (MDA), the activities of alanine aminotransferase in CAP60 and CAP45 groups, and acid phosphatase in the CAP60 group were significantly higher, while the content of total cholesterol in CAP60 and CAP45 groups was significantly lower. In the liver, compared to the control group, the content of MDA in the CAP60 group was significantly higher. 3-hydroxy-3-methylglutaryl coenzyme A reductase in CAP30 to CAP60 groups and farnesoid X receptor in CAP60 were significantly upregulated. In distal intestines, the activities of trypsin and superoxide dismutase of CAP30 to CAP60 groups were significantly lower than the FM group. In conclusion, for pearl gentian grouper, CAP could replace up to 45% of the fishmeal in the feed, while a 60% replacement level will affect cholesterol bile acid metabolism and health.

The substitution of high-level soy meals for fish meal (FM) generally leads to fish enteritis, accompanied by significant variations in gut flora. Relevant studies have pointed out a close relationship between tryptophan metabolism mediated by gut flora and vertebrate inflammatory bowel disease. Present study examines the role of tryptophan metabolism and gut flora profile in fish enteritis caused by different soybean meals. The 960 groupers were randomly assigned into 4 groups (n = 4), which including: (1) FM (the control group, fed with 50% FM feed), (2) SBM40 (replacing 40% FM with soybean meal), (3) SPC40 (replacing 40% FM with soybean protein concentrate), and (4) FSBM40 (replacing 40% FM with fermented soybean meal). Under average temperature and natural light, the groupers were cultivated with feeds of iso-nitrogen and iso-lipid for 10 weeks. The results showed that soybean meal feeds at all experimental levels had negative effects on fish gut physiology and growth performance. Typical enteritis features and fluctuations of immune system occur, which can be observed in the enzyme activities of total superoxide dismutase and lysozyme and in the contents of immunoglobulin M, complement 3 and complement 4. 16SrDNA high-throughput sequencing indicated that it greatly influenced the gut flora with the abundance of maleficent bacteria, like Vibrio, amplified with increasing dietary soybean meals. According to the “3 + 2” full-length transcriptome sequencing, soy meals at the three experimental levels inhibited the key gene expressions of tryptophan metabolic pathway in fish gut, however, there are some differences in the types of key genes that are inhibited. The canonical correlation analysis showed that the changes in key gene expressions in tryptophan metabolic pathway had a positive correlation with the expressions of pro-inflammatory genes (P < 0.05) and negatively correlated with the expression of anti-inflammatory genes (P < 0.05). It is speculated from this study that tryptophan metabolism is closely related to fish soy meal-related enteritis, and the abnormal tryptophan metabolism caused by intestinal flora imbalance may play an important role. In the future research, we can further study the tolerance of fish to soy meals feed from two aspects of tryptophan metabolism and intestinal flora changes.

Fermented soybean meal and soybean protein concentrate are products of soybean that have been processed physically or biologically, and their use as an alternative to fish meal results in a significant reduction in the effects of anti-nutritional factors (ANFs) in soybean on aquatic species. Replacing fish meal with soybean protein concentrate and fermented soybean meal can meet the high protein requirements of carnivorous fish while effectively reducing aquaculture costs; however, excessive substitution can also cause economic losses. In this study, we used transcriptome sequencing to investigate the impacts of fermented soybean meal and soybean protein concentrate on the growth and physiology of pearl gentian grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂) juveniles and to examine the mechanisms by which fermented soybean meal and soybean protein concentrate impair the intestinal condition of fish. Originally weighed 12.55 ± 0.06 g, the selected pearl gentian groupers were categorised into three treatment groups: one group was fed fish meal-based diets (FM, control group), one group was fed fish meal- and soybean protein concentrate-based diets (SPC40) and one group was fed fish meal- and fermented soybean meal-based diets (FSBM40), with the same crude protein and crude fat content in all three diets. The experiment was conducted for 10 weeks. The growth results showed that both the fermented soybean meal and soybean protein concentrate diets significantly inhibited the growth of the fish. Based on the results of enzyme activity, substance content and gene expression levels associated with intestinal damage and intestinal inflammation, it is highly likely that the fermented soybean meal and soybean protein concentrate diets affected the intestinal health of the fish and triggered intestinal inflammation. This study provides a theoretical basis to further explore the mechanism of soybean-initiated intestinal problems in fish.

This study explored the role of replacing fish meal protein with fermented soybean meal (FSBM) protein on the growth performance and intestinal morphology, immunity, and microbiota of the pearl gentian grouper (Epinephelus fuscoguttatus♀ × E. lanceolatus♂). Three isonitrogenous and isolipidic diets with increasing levels of FSBM (0%, 20% and 40%; referred to as FM, FSBM20 and FSBM40 diets, respectively) as a replacement for fish meal were selected for this study. The pearl gentian grouper were fed these diets for 10 weeks. The findings revealed that the growth of fish fed the FSBM diets (FSBM20 and FSBM40) were remarkably lower than the fish fed the FM diet. Pathological manifestations of intestinal inflammation, such as shortened intestinal mucosal folds and thickened lamina propria, were observed in the fish fed the FSBM diets. Moreover, the gene expression levels of IL1β, IL12, IL17, and TNFα were remarkably upregulated in fish fed the FSBM40 diet, in contrast to the gene expression levels of IL4, IL5, IL10, and TGFβ1, which were remarkably downregulated (p < 0.05). The FSBM diets significantly affected the stability of the fish gut microbiota. Photobacterium was the dominant phylum in all experimental groups, and the proportion of these bacteria gradually decreased with increasing FSBM substitution. The composition of intestinal flora at the genus level was not the same in the three experimental groups, with a richer composition of intestinal bacteria detected in the FSBM20 and FSBM40 groups (p < 0.05). The correlation between intestinal flora balance and immune gene expression revealed that only Photobacterium was negatively correlated with the above upregulated genes, while other bacteria were positively correlated with these pro-inflammatory factors (p < 0.05). Photobacterium was positively correlated with the above downregulated genes, while other bacteria were negatively correlated with these anti-inflammatory factors (p < 0.05). In conclusion, high levels of substitution of FSBM for fish meal causes intestinal inflammation in pearl gentian grouper. This is likely associated with changes to the intestinal flora. More attention should be paid to the negative role of dietary FSBM on intestinal flora.